Advancing our knowledge of viral membrane fusion and of IDP-membrane interactions by ESR

通过 ESR 增进我们对病毒膜融合和 IDP-膜相互作用的了解

• 批准号: 10798605
• 负责人: Jack H Freed
• 金额: $ 5.39万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10798605
• 关键词: 2019-nCoV; Binding; Binding Proteins; Calcium; Cells; Complement; Disease; Docking; Ebola virus; Electron Spin Resonance Spectroscopy; Equipment; Fluoroscope; Fluoroscopy; Funding; Funding Opportunities; Glycoproteins; Grant; HIV; Invaded; Knowledge; Learning; Lipids; Magnetism; Measurement; Membrane; Membrane Fusion; Membrane Proteins; Methods; Peptides; Principal Investigator; Process; Proteins; System; Techniques; Technology; Transmembrane Domain; Vesicle; Viral; Virus; Work; experimental study; influenzavirus; novel; particle; programs

Program Director/Principal Investigator (Freed, Jack, H.): Project Summary/Abstract This is a supplementary request for funding for equipment for the MIRA grant (R35GM148272), per Funding Opportunity PA-20-272. One of the major objectives of our R35 grant is to understand the structural mechanism(s) underlying the viral membrane fusion process induced by the fusion peptide (FP) and the transmembrane domain (TMD) of glycoproteins of an enveloped virus such as SARS-CoV-2, Ebola Virus, influenza virus, and HIV. Our advanced ESR technology has revealed major structural details of the interactions between peptides and lipids in membranes, on both the peptides and membranes. However, it will be informative if we can carry out functional studies in parallel to complement our structural studies. In fact, using ESR we have detected the function of the FP in altering the structure of the membrane, which is the initial step of the membrane fusion process. However, we need a method to detect the final step of membrane fusion, in which the membranes of the two vesicles fuse together. Fluoroscopy is the ideal and well-established technique for this task. In addition, this technique can also be applied to our novel pseudo-viral particle-vesicle docking system. Our other major objective is to study Intrinsic Disordered Protein (IDP)-membrane interactions. Many such interactions are calcium dependent, including the viral FP. Thus, measurements of Ca2+-IDP binding constant and the IDP-membrane participation constant at different Ca2+ concentrations are important, so we can learn what percentage of the IDPs in our structural study is in the membrane binding condition. Fluoroscopy is a convenient technique to obtain these parameters. The results obtained from fluoroscopy can also be compared to and complement those of the parallel measurements from our ESR experiments. OMB No. 0925-0001/0002 (Rev. 03/2020 Approved Through 02/28/2023) Page Continuation Format Page

计划总监/首席研究员（Freed，Jack，H。）： 项目摘要/摘要 这是每次资金 机会PA-20-272。我们R35赠款的主要目标之一是了解结构 融合肽（FP）诱导的病毒膜融合过程的机制和 包膜病毒的糖蛋白的跨膜结构域（TMD），例如SARS-COV-2，埃博拉病毒，埃博拉病毒， 流感病毒和艾滋病毒。我们先进的ESR技术揭示了相互作用的主要结构细节 肽和膜上的肽和脂质之间的肽和脂质之间。但是，这将是有益的 如果我们可以同时进行功能研究以补充我们的结构研究。实际上，使用ESR我们有 检测FP在改变膜结构中的功能，这是 膜融合过程。但是，我们需要一种检测膜融合的最后一步的方法，其中 两个囊泡的膜融合在一起。透视镜是理想且完善的技术 任务。另外，该技术也可以应用于我们新型的伪病毒粒子对接系统。 我们的另一个主要目的是研究固有的无序蛋白（IDP） - 膜相互作用。许多这样的 相互作用取决于钙，包括病毒FP。因此，测量Ca2+-IDP结合常数 并且在不同Ca2+浓度下的IDP膜参与常数很重要，因此我们可以学习 在我们的结构研究中，IDP的百分比在膜结合条件下。荧光镜是 方便的技术以获取这些参数。也可以比较从荧光镜检查获得的结果 从我们的ESR实验中进行并补充平行测量值的和补充。 OMB No. 0925-0001/0002（Rev. 03/2020通过02/28/2023批准）页面延续格式页面